Fig. 7
Validation of TFs involved in the regulation of PD-L1 based on rs822336 allele-specificity. H1975G/G and H1299C/C cells were seeded into T75 flasks at a density of 5 × 106 cells. Following a 48 h incubation at 37 °C in a 5% CO2 atmosphere, DNA-pull down assay was performed with immobilized wt/mut oligos incubated with nuclear extracts on 4 distinct sample group combinations. Nuclear extract of H1299C/C cells was incubated with the mut oligo; nuclear extract of H1299C/C cells was incubated with the wt oligo; nuclear extract of H1975G/G cells was incubated with the mut oligo; nuclear extract of H1975G/G cells was incubated with wt oligo. Putative TFs of the rs822336 region of PD-L1 based on its allele-specificity were detected by LC-MS/MS. (A) Venn diagram represents the overlap of 315 TFs detected over the 4 analysed sample groups. (B) Pie chart shows the gene ontology (GO) classification of TFs detected over the 4 analysed sample groups. (C) Heat map summarises the average amount measured based on the observed label-free quantitation (LFQ) intensities for each of the 315 proteins detected in the 4 analysed sample groups. (D) Volcano plot shows a proteomic based comparison between H1299C/C+mut and H1975G/G+wt. Reported points indicate proteins that display both large magnitude fold-changes (x axis) and high statistical significance (y axis). Corresponding points to C/EBPβ and NFIC are highlighted